Part:BBa_K4344041
pEX-A258-HSV-UL19-6xHis
Synthesized plasmid by Eurofins contains modified UL19 sequence. Plasmid with additional His-tag for purification.
Sequence and Features
- 10INCOMPATIBLE WITH RFC[10]Illegal EcoRI site found at 1055
Illegal XbaI site found at 1043 - 12INCOMPATIBLE WITH RFC[12]Illegal EcoRI site found at 1055
Illegal NheI site found at 1049 - 21INCOMPATIBLE WITH RFC[21]Illegal EcoRI site found at 1055
Illegal XhoI site found at 3463 - 23INCOMPATIBLE WITH RFC[23]Illegal EcoRI site found at 1055
Illegal XbaI site found at 1043 - 25INCOMPATIBLE WITH RFC[25]Illegal EcoRI site found at 1055
Illegal XbaI site found at 1043
Illegal NgoMIV site found at 307
Illegal NgoMIV site found at 454
Illegal NgoMIV site found at 814 - 1000COMPATIBLE WITH RFC[1000]
The sequence of UL19 encoding for HSV-1 major capsid protein was extracted from the complete https://www.ncbi.nlm.nih.gov/nuccore/KF498959.1 human herpesvirus 1 genome. The sequence was human codon optimised using https://www.benchling.com/ Benchling. The sequence was screened for published antibody binding sites and functional structure elements using the https://www.uniprot.org/ UniProt database. We identified the amino acids 862 to 880 in UL19 as a published antibody binding site (Han et al., 2019). Since it is a published antibody site, we anticipate that the corresponding DNA sequence is highly conserved in the HSV genome. Therefore, we chose a sequence for cloning and expression including this structure element. For UL19 bases 2446 to 3444 were selected resulting in a fragment of 999 bp in size. The fragment was further modified to mask unwanted restriction sites and ease primer design while keeping the amino acid sequence unchanged. UL19 was modified at the 16-18TCC>AGT and 21G>A. A His-tag (5’-CATCACCATCACCATCAC-3’) and a TAA stop codon were added at the 3’ sequence end.
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